This application under the Academic Research Enhancement Award (AREA) program proposes research which would endeavor to develop modern methods for the differentiation of species and populations of mosquito which breed in treeholes and other containers both natural and man-made. The project stresses the development of new morphometric methods based on mathematical shape analysis for identifying the ova of the aedine species in question. An automated system for data acquisition and analysis is also proposed for development. This project has three long-term goals. The first of these is to develop new, unique morphometric characters associated with the egg stage which can be used to identify species of Aedes which utilize treeholes and containers for oviposition and larval development. In order to test the hypothesis that species can be delineated using egg shape the investigators will first endeavor to develop an automated system for routine collection and analysis of data on egg size and shape and chorionic sculpturing. Technical aspects of this task will be accomplished by employing a charge coupled device (CCD) to send microscopic images of eggs to a frame grabber, digitizing tablet and computer. Fourier and eigenshape analysis will be conducted using specialized software, and statistical comparisons of the resulting parameters between groups will be made. To be scrutinized by these methods are Aedes triseriatus, Ae. hendersoni, and Ae. albopictus, species presently maintained in the applicant's laboratory, and Ae. atropalpus, Ae. aegypti and Ae. bahamensis for which starting material will be provided by collaborators. Eggs for analysis will be obtained from individual bloodfed females. In addition to interspecific studies, the aforementioned methods will be applied to the separation of geographic populations within species based on egg size and shape. These studies will test the hypotheses that geographic variation can be demonstrated in egg shape parameters and that the relatedness of populations can be deduced from these data. Various geographic strains of Aedes triseriatus, Ae. hendersoni and Ae. albopictus are available in the applicant's laboratory for these studies. The second long term goal of the project is to provide the basis for further research beyond this project. This work will utilize the automated system to investigate chorionic sculpturing patterns as possible bases for species and population identification. The relationship of egg shape and size to geographic location, environmental conditions and seasonality will be studied. Finally, the effect of hybridization on egg shape variability will be investigated. The final large objective of the proposal is to train undergraduate students in mathematically and statistically based morphological systematics. The progress report lists numerous students from the applicant's program who have successfully completed research projects and who have won student research competitions sponsored by national scientific societies.